Juxtaglomerular Cells Grown as Monolayer Cell Culture Contain Renin, Angiotensin I‐Converting Enzyme, and Angiotensins I and II/III
Juxtaglomerular Cells Grown as Monolayer Cell Culture Contain Renin, Angiotensin I‐Converting Enzyme, and Angiotensins I and II/III
复制标题
作为单层细胞培养物生长的肾小球旁细胞含有肾素、血管紧张素 I 转换酶以及血管紧张素 I 和 II/III
作者：
W. Rightsel;T. Okamura;T. Inagami;J. Pitcock;Y. Takii;B. Brooks;P. Brown;E. Muirhead
采用新生大鼠肾皮质细胞单层培养法，培养肾小球细胞（JGC）。JGC具有肾脏内的特征，包括表明平滑肌起源的外周致密体和肌原纤维;含有与蛋白质合成一致的蓬松物质的粗糙ER;用于包装颗粒的突出高尔基体，以及具有分泌颗粒和溶酶体特征的颗粒。将培养的细胞移植到同基因受体中，存活2周或更长时间，并保留JGC的特征。当用兔抗纯大鼠肾素抗体和异硫氰酸荧光素标记的羊抗兔IgG重链特异性片段F（ab′）2（Fc片段）处理JGC颗粒时，JGC颗粒发出荧光。后者表明存在肾素。在DFP、卡托普利、亮抑酶肽和EDTA存在下裂解JGC，并在胃蛋白酶抑制剂存在下提取。溶胞产物中含有被特异性肾素抗体抑制的肾素活性。非特异性蛋白酶被抗体及其最适pH排除。在不使用EDTA和卡托普利的情况下制备的裂解物中检测到血管紧张素I转化酶。血管紧张素I和11/ III来自提取物通过额外的提取，TLC和RIA，使用高度特异性的抗体。血管紧张素通过色谱法以真实的血管紧张素监测来确认。我们的结论是，培养的JGC含有肾素，血管紧张素I转换酶，血管紧张素I和II/III。
A monolayer cell culture of juxtaglomerular cells (JGC) was derived from the renal cortex of neonatal rats. The JGC had the characteristics of those within the kidney, including peripheral dense bodies and myofibrils indicating a smooth muscle origin; rough ER containing fluffy material consistent with protein synthesis; a prominent Golgi apparatus for packaging granules, and granules having the characteristics of secretory granules and lysosomes. Transplants of the cultured cells into syngeneic recipients survived for 2 weeks or longer and retained the features of JGC. The JGC granules fluoresced when treated with a rabbit antibody against pure rat renin, followed by fluorescein isothyocyanate conjugated F(ab′)2 fragment of goat antirabbit IgG (Fcfragment) heavy chain specific. The latter indicated the presence of renin. The JGC were lysed in the presence of DFP, captopril, leupeptin, and EDTA, and were extracted in the presence of pepstatin. The lysate contained renin activity that was inhibited by a specific renin antibody. Nonspecific proteases were excluded by the antibody and its pH optimum. Angiotensin I-converting enzyme was detected in the lysate prepared without the use of EDTA and captopril. Angiotensins I and 11/ III were derived from the extract by additional extractions, TLC, and RIA, using highly specific antibodies. The angiotensins were confirmed by chromatography monitored by authentic angiotensins. We concluded that the cultured JGC contained renin, angiotensin I-converting enzyme, and angiotensins I and II/III.
血管紧张素 II 免疫反应性与肾素共存于肾脏的肾小球旁颗粒细胞中。
DOI：
10.1073/pnas.78.6.3897
发表时间：
1981
影响因子：
11.1
作者：
Celio,MR;Inagami,T
通讯作者：
Inagami,T